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Tyrosine derived polycarbonate membrane is useful for guided bone regeneration in rabbit mandibular defects.

Standardized bilateral through-and-through defects (12x6 mm) were created extraorally in the mandibular angle of 18 New Zealand White rabbits. Animals were divided in to three groups (n=6) according to the intended healing time. On the left side, defects were covered with a poly(desaminotyrosyl-tyrosine-ethyl ester carbonate) (PDTE carbonate) membrane wrapped around the inferior border of the mandible and fixed with bioabsorbable sutures. On the right side, the defects were filled with a mesh made of bioactive glass 13-93 and 3 wt% chitosan. The defects were covered with the same membranes. Periosteal flap was sutured over the membrane. Radiographically, bone ingrowth was seen in all specimens at 12 weeks postoperatively. At 24 weeks, completely ossified area remained approximately at the same level as at 12 weeks, but the non-ossified area decreased to almost zero. However, the bioactive glass mesh did not improve the results. Nevertheless, enveloping the defect with PDTE carbonate membrane seemed to play a crucial role in new bone formation. Based on these results, we conclude that tyrosine polycarbonate is a promising new material for guided bone regeneration.

Animals↗

Treatment of ligature-induced peri-implantitis by lethal photosensitization and guided bone regeneration: a preliminary histologic study in dogs.

BACKGROUND: The purpose of this pilot study was to evaluate the healing potential and reosseointegration in ligature-induced peri-implantitis defects adjacent to various dental implant surfaces following lethal photosensitization. METHODS: A total of 36 dental implants with 4 different surface coatings (9 commercially pure titanium surface [CPTi]; 9 titanium plasma-sprayed [TPS]; 9 hydroxyapatite [HA]; and 9 acid-etched [AE]) were inserted in 6 male mongrel dogs 3 months after extraction of mandibular premolars. After a 2-month period of ligature-induced peri-implantitis and 12 months of natural peri-implantitis progression, only 19 dental implants remained. The dogs underwent surgical debridement of the remaining dental implant sites and lethal photosensitization by combination of toluidine blue O (100 microg/ml) and irradiation with diode laser. All exposed dental implant surfaces and bone craters were meticulously cleaned by mechanical means, submitted to photodynamic therapy, and guided bone regeneration (GBR) using expanded polytetrafluoroethylene (ePTFE) membranes. Five months later, biopsies of the implant sites were dissected and prepared for ground sectioning and analysis. RESULTS: The percentage of bone fill was HA: 48.28 +/- 15.00; TPS: 39.54 +/- 12.34; AE: 26.88 +/- 22.16; and CPTi: 26.70 +/- 16.50. The percentage of reosseointegration was TPS: 25.25 +/- 11.96; CPTi: 24.91 +/- 17.78; AE: 17.30 +/- 15.41; and HA: 15.83 +/- 9.64. CONCLUSION: These data suggest that lethal photosensitization may have potential in the treatment of peri-implantitis.

Acid Etching, Dental↗

Bonelike/PLGA hybrid materials for bone regeneration: preparation route and physicochemical characterisation.

Bonelike/PLGA hybrid materials have been developed using gamma-MPS as silane-coupling agent between the inorganic and organic phases for controlled drug delivery applications. Silanization showed to be more effective when cyclohexane was used as a non-polar solvent (nP method) due to a chemical interaction between Bonelike, and the silane film, while by using a 95/5 (V/V) methanol/water as a polar solvent (P method), a much thinner film was achieved. Functional groups of PLGA, such as the carbonyl group (C=O), were identified using Raman and FTIR-ATR analysis and therefore these groups may be used to link therapeutic molecules. These novel hybrid materials prepared by combining silanization and post-hybridisation processes are expected to find use in medical applications of bone regeneration and as drug delivery carrier for therapeutic molecules.

Bone Regeneration↗

Autologous concentrated platelet-rich plasma (cPRP) for local application in bone regeneration.

Platelets are rich in growth factors that may contribute to an accelerated tissue regeneration process. The therapeutic osteogenic effect of local platelet administration probably depends on the amount of growth factors delivered within. To improve platelet-derived factor preparations, the platelets have to be concentrated without loss of the granular growth factor load. An autologous procedure according to the Good Manufacture Practice (GMP) guidelines to prepare a high concentrate from platelet-rich plasma (cPRP) for clinical application in bone regeneration is presented. Platelet-rich plasma (PRP) was generated from the centrifugation supernatant of 250 ml anticoagulated autologous blood. The PRP was then further centrifuged at the day of operation to generate concentrated PRP (cPRP) in 8 ml plasma. The data from 34 patients (9 m, 25 f), ageing from 21 to 71 years (median 50.4) were evaluated. Both, the platelet counts as well as the TGF-beta1 concentrations in the cPRP were found to be 12-fold increased compared to the PRP values. Overall, a 17-fold increase in platelet concentrations was achieved compared to the patients' whole bloods and platelet counts correlated with increased levels of TGF-beta1 within the cPRP. These results give rise to quality control specifications for a safe and cost effective preparation of cPRP. The method has proven to be suitable for blood services to support the needs for treatment in oral and maxillofacial surgery.

Adult↗

Combined implantation of particulate dentine, plaster of Paris, and a bone xenograft (Bio-Oss) for bone regeneration in rats.

UNLABELLED: The purpose of this study is to assess the combination of particulate dentine and plaster as bone substitute material in calvarial bone defects in rats, and to compare it with a bone xenograft (Bio-Oss). MATERIAL AND METHODS: Forty rats were assigned randomly to five groups and each group was further divided into two subgroups, 8 and 16 weeks after implantation. The defect was filled with different graft materials in each group: Group 1, defects were filled with particulate dentine and plaster using a 2:1 ratio; Group 2, defects were filled with particulate dentine; plaster, and Bio-Oss using a 2:1:1 ratio; Group 3, defects were filled with plaster and Bio-Oss using a 1:1 ratio; Group 4, defects were filled with Bio-Oss only; and Group 5, untreated control defects. Histological sections and histomorphometric analysis of defects were obtained at 8 and 16 weeks postoperatively. RESULTS: New bone formation was highest in Group 4, followed by Group 3, than Group 2, Group 1, and finally the control group. CONCLUSION: The combination of particulate dentine and plaster is an alternative bone substitute, although it is less effective than Bio-Oss.

Analysis of Variance↗

A comparative study of anorganic xenogenic bone and autogenous bone implants for bone regeneration in rabbits.

The aim of this study was to investigate the possible use and ultimate fate of anorganic xenogenic bone for the restoration of defects in the maxillae and mandibles of adult rabbits. Furthermore, anorganic xenogenic bone was compared with implanted autogenous bone particles with regard to the response of the surrounding connective tissue and possible resorption of these implants by multinucleated cells. Results showed that after 12 weeks, the implanted autogenous bone was actively resorbed by multinucleated cells, and new bone was formed in close apposition to the particles. In contrast, implanted anorganic xenogenic bone was degraded to a much lesser extent, and new bone was seen adjacent to the anorganic bone particles without signs of resorption. Further long-term studies are needed to determine whether anorganic xenogenic bone may be regarded as a resorbable material and whether any side effects occur as a result of this material's tendency to linger on in the recipient bed.

Animals↗

Long-term results of implants treated with guided bone regeneration: a 5-year prospective study.

The aim of this prospective 5-year longitudinal study was to follow endosteal implants in which guided bone regeneration (GBR) was applied during implant placement. In 75 patients, defects around implants (Branemark System) were treated with Bio-Oss and Bio-Gide (112 implants). In split-mouth patients in this group, Bio-Oss and Gore-Tex were used in the second defect site (41 implants). All 75 patients had at least 1 implant that was entirely surrounded by bone and served as the control (112 implants). After placement of the definitive prostheses (single-tooth, fixed, or removable implant prostheses), patients were recalled after 6 months and then every 12 months during a 5-year observation period. The following variables were investigated: implant survival, marginal bone level (MBL), presence of plaque, peri-implant mucosal conditions, height of keratinized mucosa (KM), and marginal soft tissue level (MSTL). The cumulative implant survival rate after 5 years varied between 93% and 97% for implants treated with or without GBR. The mean MBL after 60 months was 1.83 mm for sites treated with Bio-Oss and Bio-Gide, 2.21 mm for sites treated with Bio-Oss and Gore-Tex, and 1.73 mm for the control sites. The MBL values were found to increase significantly with time and differed significantly among the treatment groups. During the observation period, KM varied between 3.16 and 3.02 mm. A slight recession of 0.1 mm was observed, and plaque was found in 15% of all sites and was associated with inflammatory symptoms of the peri-implant mucosa. It was observed that such symptoms and recession correlated more strongly with the type of restoration than with the type of treatment. This study demonstrated that implants placed with or without GBR techniques had similar survival rates after 5 years, but that bone resorption was more pronounced in sites with GBR treatment. It was assumed that the use of GBR is indeed indicated when the initial defect size is larger than 2 mm in the vertical dimension.

Adult↗

Bone marrow stromal cells and their use in regenerating bone.

Tissue engineering approaches have recently been devised to repair large bone losses. Tissue engineering takes advantages of the combined use of cultured living cells and 3D scaffolds to deliver vital cells to the damaged site of the patient. Cultured bone marrow stromal cells (BMSCs) can be regarded as a mesenchymal progenitor/precursor cell population derived from adult stem cells. When implanted in immunodeficient mice, BMSCs combined with mineralized 3D scaffolds to form a primary bone tissue that is highly vascularized. We have used autologous BMSC/bioceramic composites to treat full-thickness gaps of tibial diaphysis in sheep. The healing process has been investigated. The sequence of events is as follows: (1) bone formation on the outer surface of the implant; (2) bone formation in the inner cylinder canal; (3) formation of fissures and cracks in the implant body; (4) bone formation in the bioceramic pores. Similar composites whose size and shape reflected each bone defect have been implanted at the lesion sites of three patients. External fixation was used. Patients have been followed for more than three years. The results obtained are very promising and we propose the use of culture-expanded osteoprogenitor cells in conjunction with hydroxyapatite bioceramics as a significant improvement in the repair of critical size long bone defects.

Adipocytes↗

Periodontal repair in dogs: expanded polytetrafluoroethylene barrier membranes support wound stabilization and enhance bone regeneration.

A wound stabilizing effect of expanded polytetrafluoroethylene (ePTFE) membranes was evaluated in supra-alveolar periodontal defects in 5 beagle dogs. The defects, 5 to 6 mm in height, were surgically created around the 2nd, 3rd, and 4th mandibular premolar teeth in contralateral jaw quadrants. The root surfaces were conditioned with heparin, which, in this model, has been demonstrated to compromise periodontal healing and result in formation of a long junctional epithelium. Wound closure included application of ePTFE membranes around each premolar tooth in one jaw quadrant in each dog and flap positioning coronal to the cemento-enamel junction in both jaw quadrants. Healing progressed uneventfully except for 3 teeth in 2 dogs, which experienced membrane exposure. The dogs were sacrificed after a 4-week healing period and tissue blocks were prepared for histometric analysis. Connective tissue repair in heparin+membrane-treated teeth averaged 98% of the defect height compared to 84% in control heparin-treated teeth (P < or = 0.05). Junctional epithelium formation was smaller in membrane-treated teeth than in control teeth (P < or = 0.05) and was usually terminated coronal to the membrane. Bone regeneration was enhanced in membrane-treated teeth compared to controls (P < or = 0.01) and was strongly correlated to the area under the membrane in teeth without membrane exposure (r2 = 0.993; P = 0.002). This correlation was reduced when teeth with membrane exposure were included in the analysis (P < or = 0.05). Cementum regeneration was minimal under both treatment conditions. Root resorption was increased in membrane-treated compared to control teeth (P < or = 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Alveolar Process↗

Wnt signaling activation during bone regeneration and the role of Dishevelled in chondrocyte proliferation and differentiation.

Wnt signaling is intrinsically involved in diverse cellular activities during cell differentiation, early embryonic development and organogenesis. Although much is known regarding the effects of Wnt signaling in the developing skeletal system, its role during regeneration remains unclear. Herein, we show transcriptional activation of specific members and target genes of the Wnt signaling pathway. Specifically, all of the Wnt signaling members and target genes analyzed were found to be upregulated during the early stages of fracture repair, with the exception of LEF1 whose expression was downregulated. In addition, spatial expression analysis of Dishevelled (Dvl) and beta-catenin in the fracture callus revealed an identical pattern of expression with both proteins localizing in osteoprogenitor cells of the periosteum, osteoblasts and proliferating/pre-hypertrophic chondrocytes. Further, in vitro knockdown of all three Dvl isoforms in chondrocytes using small interfering RNAs (siRNA) leads to partial inhibition of cell proliferation and differentiation, decreased expression of chondrogenic markers (ColII, ColX, Sox9) and suppressed nuclear accumulation of unphosphorylated beta-catenin. Taken together, these data verify our previous finding that the Wnt signaling pathway is activated during bone regeneration, by characterizing the temporal and spatial expression of a broad spectrum of Wnt-signaling molecules. Our data also suggest that all three Dvl isoforms, acting through the Wnt canonical pathway, are critical regulatory molecules for chondrocyte proliferation and differentiation.

Adaptor Proteins, Signal Transducing↗

Tissue-engineered injectable bone regeneration for osseointegrated dental implants.

The present study investigated a correlation between osseointegration in dental implants and an injectable tissue-engineered bone, using mesenchymal stem cells (MSCs) and platelet-rich plasma (PRP). Initially, the teeth in the mandible region were extracted and the healing period was 1 month. Bone defects on both sides of the mandible were prepared with a trephine bar. The defects were implanted with graft materials as follows: PRP, dog MSCs (dMSCs), and PRP, autogenous particulate cancellous bone and marrow (PCBM), and control (defect only). Two months later, the animals were evaluated by histology, and at the same time dental implants were installed. Two months later, the animals were sacrificed and nondecalcified sections were evaluated histologically and histometrically. According to the histological observations, the dMSCs/PRP group had well-formed mature bone and neovascularization, compared with the control (defect only) and PRP groups, as was the same for the PCBM group. A higher marginal bone level was observed around implants with PRP, PCBM, and dMSCs/PRP compared with the control. Furthermore, the values describing the amount of bone-implant contact (BIC) at the bone/implant interface were significantly different between the PRP, PCBM, dMSCs/PRP, and control groups. Significant differences were also found between the dMSCs/PRP and control groups in bone density. The findings of this experimental study indicate that the use of a mixture of dMSCs/PRP results in good results such as the amount of BIC and bone density comparable with that achieved by PCBM.

Animals↗

Guided bone regeneration of cranial defects, using biodegradable barriers: an experimental pilot study in the rabbit.

The aim of this study was to test if a biodegradable barrier could be used to achieve proper bone healing of full-thickness trephine skull defects, applying the biological principle of guided tissue regeneration (GTR). Two New Zealand white rabbits were used. In each animal, 2 circular through-and-through bone defects with a diameter of 8 mm were created in the midline of the frontal and parietal bones of the calvarium. One defect was covered with the mucoperiosteal flaps without placement of an intervening membrane barrier (control). One test defect (test 1) was covered by a biodegradable, non-porous polylactic acid membrane on the outer (supra-calvarial) side of the defect, and 2 test defects (tests 2 and 3) were covered by similar membranes on both the outer and the inner aspects of the defects, prior to flap closure. 6 weeks postsurgically, the animals were sacrificed and the defect areas including surrounding tissues were harvested for histological preparation. The control defect was essentially occupied by supra-calvarial soft tissue, located in direct contact with the dural tissue. In the test cavities, there was a continuous bridge of regenerated bone extending from one edge of the defect to the other, although in test 1 not attaining the same thickness as the bone bordering the defect. In the 2 other test defects, the regenerated bone had reached a thickness almost corresponding to that of the surrounding bone. The bone regeneration was achieved without recourse to adjunctive bone graft materials.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Histological aspects in bone regeneration of an association with porous hydroxyapatite and bone marrow cells.

The osteogenic potential of an association of two kinds of hydroxyapatite (HA) porous ceramics with autologous bone marrow cells cultured with or without dexamethasone (10(-8)M) addition in the culture medium and non-cultured rabbit marrow stromal stem cells (MSCs) was tested after 4 weeks of implantation in the dorsal muscles of spine in rabbit. A significantly higher number of rabbits with implants containing bone tissue inside pores were obtained with 10(7) cells ml(-1) cultured treated with Dex. In the HA porous ceramic using naphtalen as porogen agent, the bone recolonization remains only at the periphery of implants and in the second row of pores, while in the HA porous ceramic using polymethacrylate (PMMA) microbeads as porogen agent, the bone recolonization is observed in the depth of implants. In the PMMA HA group, the Krüskal-Wallis variance analysis between the rabbits is significantly different with the percentage of number of occupied pores and occupied pores with bone tissue is different (p<0.05).

Journal Article↗

Bone regeneration with resorbable polymeric membranes: treatment of diaphyseal bone defects in the rabbit radius with poly(L-lactide) membrane. A pilot study.

Tubular poly(L-lactide) membranes with a pore size of 5-15 microns and a molecular weight of 70,000 Daltons were implanted into 24 New Zealand skeletally mature rabbits to cover 10-mm mid-diaphyseal defects of the radius of the forelimb. An identical defect on the contralateral limb was not treated with the membrane and served as a control. The animals were killed at 1, 2, 4, 8, 12, 24, 36, and 64 weeks after implantation, and radiographic and microscopic studies were conducted. The canals of the polymeric tubes were initially filled with blood. At 2 weeks, there was direct woven bone formation within the polymeric tube in continuity with the fragment cortices and its medullary canal. The formation of woven bone across the defect progressed until reconstruction of the defect had occurred at 6-8 weeks. The bone continued to remodel throughout the observation period of 64 weeks. By 12 weeks, bone within the lumen of the implant consisted of cancellous bone and cortical bone lining the membrane walls. At 24, 36, and 64 weeks, the implants were filled with cancellous bone and cortical bone in direct apposition to the polymer membrane. For one implant, the newly formed woven bone had only incompletely filled the defect at 8 weeks. This resulted in a nonunion with a residual gap of 0.5 mm and the appearance of mature bone. There was extensive bone formation along the intraosseous membrane in both control and implanted defects, although the untreated defects were rapidly filled with overlying muscle and soft tissues. The osseous activity of the untreated defects appeared confined to the bone ends by the interposed muscle and fibrous soft-tissue margins. The untreated defects resulted in radial-ulnar synostosis along the intraosseous membrane with cortical bone caps at the bone ends. The poly(L-lactide) membrane remained intact throughout the 64-week period without evidence of significant degradation. The membranes were encapsulated in a thin fibrous tissue. There was no histological evidence of acute or chronic inflammation associated with the implants.

Animals↗